LOCUS       sequence-13220-e        4044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4044)
  AUTHORS   Yilmaz A, Nishiyama MY Jr, Fuentes BG, Souza GM, Janies D, Gray J, 
            Grotewold E
  TITLE     GRASSIUS: a platform for comparative regulatory genomics across the 
            grasses.
  JOURNAL   Plant Physiol. 2009 Jan;149(1):171-80. doi: 10.1104/pp.108.128579. 
            Epub 2008 Nov 5.
  PUBMED    18987217
REFERENCE   2  (bases 1 to 4044)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1104/pp.108.128579"; journalName: "Plant Physiol"; date: 
            "2009-01"; volume: "149"; issue: "1"; pages: "171-80"
COMMENT     Sequence Label: pUT1673-HQ858864
FEATURES             Location/Qualifiers
     source          1..4044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             112..132
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1601..1700)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1717..1736)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1718..1736)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1741..1757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1741..1757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1859..1878)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1870..2679
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1937..1956)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2826..3414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3315..3334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3568..3585
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3744..3771)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3863..3949)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3998..4020
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4012..4029
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4013..4029
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcggccgc cttgtttaac
      121 tttaagaagg agcccttcac catgaactgc gggccgcccg accagttgcc gccggcgacg
      181 gcgccgtcgt gcttcctcaa cctcaactgg gaccagtcca tggacgcggc ggcgggaggg
      241 cacctcgacc cggcgctcag ctccatggtg tcctcgccgg cgtccaactc gacgggcgct
      301 ctccacggga tctcgccgca gccgcactac ggtggcggga cgccgctcag ctcgcccccc
      361 aagctcaacc tgtccatgat ggggcagttc caccactacg ccgcgccgcc gcaggtgggc
      421 ggcggcggag gcggaggcgg gggcctgcca atcctcgaga acctgatgcc catgggccac
      481 ctcgaccagt tcctcgccga ccctggcttc gccgagcgcg ccgcgaggct ctccggcttc
      541 gacgcccgcg gcggcggcgg aggaggaggc tacggcggcg ccggcccggc gcaattcggc
      601 ctccctgacg ccggcgcggc cggcgcatcg aaggagatgg agctcgggaa cacccgggac
      661 gagtcgtcgg tgtctgatcc ggcgcccggc ggcgccgaga ttccgcccaa gggggcttcc
      721 gacggcaatg cacggaagcg gaaggcctcc gggaagggca aaggcaagga cagccccatg
      781 tccacctccg ccgccaagga ggattccagc ggcaagcgtt gcaaatcgac ggaggagagc
      841 aatgcggccg ccgaggagaa ttccggcaag gggaaggccg cgcagagcaa cagcgagaat
      901 ggcggcggca agaagcaagg gaaggacagc tcgtcgaagc cccccgagcc gcccaaggac
      961 tacatccatg tccgcgcccg gcgcggcgag gcgacggaca gccacagcct cgccgagagg
     1021 gtgagaagag agaagataag ccagaggatg aagctgctgc aggatctcgt gccgggttgt
     1081 aacaaggtgg ttggaaaggc tgtcatgctc gatgaaatca taaactatgt gcagtcgttg
     1141 caacggcaag tcgagttttt gtccatgaag ttggccactg tgaatcccca gctggacttc
     1201 aacaatttgc ctaacctcct tgctaaagat atgcaccagt catgcagccc gttacagagc
     1261 tcacattttc cactagagac ctcaggtgcg ccgctgccct acattaacca gcctcagcaa
     1321 gggaaccctc tcggttgtgg cctgacgaac ggcatggaca accagggttc tatgcaccca
     1381 ttggacccgg cattttgccg gccaatgggc tcgcaccatc ctttcctcaa tggggttagc
     1441 gatgcggctt ctcaggttgg tgctttctgg caagatgatc tccaaagtgt tgttcagatg
     1501 gatatggggc aaagtcagga gatcgccacc tcttccaata gctacaatgg atcgttgcaa
     1561 acagtccaca tgaaaatgga gcttaagggt gggcgcgccg acccagcttt cttgtacaaa
     1621 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1681 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     1741 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1801 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1861 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1921 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1981 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2041 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2101 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2161 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2221 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2281 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2341 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2401 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2461 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2521 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2581 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2641 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2701 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2761 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2821 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2881 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2941 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3001 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3061 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3121 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3181 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     3241 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3301 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3361 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3421 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3481 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3541 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3601 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3661 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     3721 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     3781 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     3841 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     3901 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     3961 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     4021 acggccagtc ttaagctcgg gccc
//
